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PCR Reagents Kits on ZAGENO

Polymerase chain reaction (PCR) is a useful molecular biology technique developed by Kary Mullis in the 1980s. PCR is used to amplify DNA templates into millions of copies of a particular DNA sequence within hours. Due to its powerful ability, PCR is applied in different areas such as; genetic and infectious disease identification, forensics, and research in molecular biology. In contrast to conventional PCR, quantitative reverse transcriptase PCR (qRT-PCR) allows measurement of gene expression quantitatively and in real-time.

In general, a complete PCR reaction requires five basic PCR reagents; DNA/RNA template, DNA polymerase, primers (forward and reverse), deoxynucleotide triphosphates (dNTPs) and PCR buffers.

Expand your understanding in our Knowledge Section. Perfect your experiments with our Troubleshooting Guides. Join in the discussion within our Community. Learn about ZAGENO at How It Works.

PCR templates

Selecting the right templates for PCR is vital as different templates are used for the various methods of PCR. For instance, when performing RT-qPCR, RNA templates are required for producing complementary DNA while DNA templates are needed for conventional PCR. It is also important to have DNA or RNA templates in high quality and quantity as this can help optimizing the efficiency of PCR. For this reason, a good PCR template preparation kit is needed for the first step of a successful PCR.

DNA polymerase

All PCR reactions require a DNA polymerase which can work under high temperature (around 70 °C) as the first phase of PCR involves separation of DNA strands at high temperature (~90 °C). Taq polymerase, which is a heat-stable enzyme isolated from the thermophilic bacterium Thermus aquaticus is a commonly used DNA polymerase for PCR. Some DNA polymerases are engineered to be activated at high temperature only, so as to reduce non-specific amplification at the beginning stage of PCR.

Primers

The initiation of DNA synthesis requires primers; short strands of nucleotides (DNA or RNA) which are complementary to the template DNA and serve as a DNA synthesis starting point for the DNA/RNA polymerase. Annealing of primers to single strand DNA requires lower temperature (50-65 °C) than the denaturation step. Once the annealing step is completed, hydrogen bonds will form between the primers and the template DNA.

Deoxynucleotide triphosphates (dNTPs)

Deoxynucleotide triphosphates (dNTPs) are the essential components of PCR as they act as the building blocks of nucleic acids; DNA polymerase cannot synthesize DNA without a supply of dNTPs.

PCR buffers

PCR buffers ensure that the PCR reaction is conducted under optimal conditions. The major components of PCR buffer include Tris-HCl, potassium chloride (KCl) and magnesium chloride (MgCl2). Tris-HCl and KCl are responsible for maintaining a stable pH during PCR. Magnesium ions act as cofactors for DNA polymerase so as to ensure proper DNA synthesis function of the polymerase during PCR. PCR buffer is usually available at 10X concentration.

Compare PCR Reagents!

With our compare function, you can avoid all the time and energy wasted sifting through multiple web pages from different suppliers. At ZAGENO you can clearly see kits side-by-side, with the relevant attributes for each kit neatly in line for easy selection of the best product for you.

Click on the comparison below for a clearer view!

This example highlights how every kit has positive and negative aspects, and that only side-by-side comparison can allow you to correctly judge which is the right kit for you.

For instance, this comparison shows:

  • that the Saveview Classic Kit from Applied Biological Sciences has the cheapest price per preparation
  • that the Precision Blue Real-Time PCR Dye from Bio-Rad has a 200X concentration.
  • that the CYGREEN Nucleic Acid Dye can also be used to stain gels.

The ZAGENO comparison engine does not show the best kit, but instead helps you, the researcher, choose the most sutiable kit for your experiment.

Go to our How it Works page for a guide to using the ZAGENO comparison engine.

Video Credits

Николай Чабан/YouTube

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